In collaboration with NIH-CRM, we previously generated iPSCs derived from neuronal precursors differentiated from the H1 (WA01) hES cell line and compared their gene expression and methylation profiles to the parental line. This research was published in 2014. We also published several papers this year on the culture of human pluripotent stem cells and are co-authors on a publication describing differentiation of iPSCs toward bone and cartilage. In collaboration with NIH-CRM, 5 transgenic hESC lines, which express traceable markers from cell type-specific promoters, were generated using a Zinc Finger nuclease-assisted gene-targeting method to integrate the transgenes into AAVS1, one of the known safe harbor sites in the human genome. Each transgene contains a gene for ZS green and a drug-selection marker. Correct transgene integration as well as normal karyotype was confirmed for each line and expression of ZS green from appropriate cell types has been confirmed in 3 lines. All lines have been deposited with WiCell and, upon ratification of appropriate agreements, will be available for distribution to the community. This manuscript is currently submitted for review. In terms of bringing pluripotent stem cells to the clinic, we have been evaluating novel xeno-free substrates, media and small molecule inhibitors as well as non-integrating methods of reprogramming. These methods include episomal plasmids, Sendai virus and microRNA boosted mRNA- based reprogramming. A novel self-replicating synthetic RNA reprogramming system will also be tested. We have extended our studies on the novel non-colony type (NCM) monolayer method for pluripotent cell culture using these different small molecules, media and alternative substrates. We will continue to improve the method for application to high thoughput and scalability for drug screening and therapeutic use. Finally, we have been involved in mentoring and teaching standard and feeder-free, pluripotent stem cell culture, assisting and advising on the generation of iPSCs from collaborators samples as well as assisting and advising on differentiation strategies as requested. As always, we update the SCU website with protocols and information as it becomes available to aid other researchers in their studies.